Luca Menabuoni

Photothermal effects in connective tissues mediated by laser-activated gold nanorods

We report a study on the application of laser-activated nanoparticles in the direct welding of connective tissues, which may become a valuable technology in biomedicine. We use colloidal gold nanorods as new near-infrared chromophores to mediate functional photothermal effects in the eye lens capsules. Samples obtained ex vivo from porcine eyes are treated to simulate heterotransplants with 810-nm diode laser radiation in association with a stain of gold nanorods of aspect ratio approximately 4. This stain is applied at the interface between a patch of capsule from a donor eye and the capsule of a recipient eye. Then, by administration of laser pulses of 40 msec and approximately 100-140 J/cm(2), we achieved the local denaturation of the endogenous collagen filaments, which reveals that the treated area reached temperatures above 50 degrees C. The thermal damage is confined within 50-70 mum in a radial distance from the irradiated area.

Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea

Corneal laser welding is a technique used clinically to induce the immediate sealing of corneal wounds. We present an experimental and model analysis of the temperature dynamics during diode laser-induced corneal welding, which is aimed at characterizing the mechanism of tissue fusion. Ex vivo tests were performed on porcine eyes in the typical irradiation conditions used for laser-induced suturing in cornea transplant. Three laser power densities (12.5 W/cm(2), 16.7 W/cm(2), 20.8 W/cm(2)) were tested. The superficial temperature of the cornea was measured by means of an infrared thermocamera. Experimental data were compared with the results of a three-dimensional (3D) model of a laser-welding process in the cornea, solved by the use of the Finite Element Method (FEM). The model solution and experimental results showed good agreement. The model was thus used to estimate the temperature enhancement inside the corneal wound and to calculate the thermal damage inside the tissue. The results indicated a selective, spatially confined heating effect that occurred at operative temperatures (59 to 66 degrees C) close to intermediate denaturation points of the stromal collagen, before its complete disorganization. No significant heat damage to the region of the laser-treated wound was evidenced in the operative irradiation conditions of corneal welding.

Laser-assisted corneal welding in cataract surgery: Retrospective study

PURPOSE: To evaluate the efficacy of laser welding to close corneal wounds in cataract surgery. SETTING: Department of Ophthalmology, Hospital of Misericordia e Dolce, Prato, Italy. METHODS: This retrospective nonrandomized interventional case series included 20 eyes of 20 patients. All eyes had cataract with visual acuity worse than 20/200. Ten patients had phacoemulsification surgery, and 10 had extracapsular cataract extraction (ECCE). Surgeries were followed by laser‐assisted closure of the corneal wounds by diode laser welding of the stroma using a technique established in animal models. Preoperative and postoperative (1, 3, 7, 30, and 90 days) astigmatism, level of inflammation, Seidel testing, and endothelial cell counts were obtained. RESULTS: There were minor differences in preoperative and postoperative astigmatism except in 1 eye in the ECCE group. The Seidel test showed no wound leakage during the follow‐up examinations, and endothelial cell loss was similar to that published for standard surgical cataract procedures. No collateral effects of the laser welding were detected, nor was postoperative inflammation. CONCLUSIONS: Astigmatism was slightly affected by the use of laser welding for corneal wound closure after phacoemulsification and ECCE. The laser‐assisted corneal sealing procedure was rapid and safe and could serve as an alternative to corneal suturing, with significant potential applications for the closure of longer incisions, such as those used in penetrating keratoplasty.

Experimental study on the healing process following laser welding of the cornea

An experimental study evaluating the application of laser welding of the cornea and the subsequent healing process is presented. The welding of corneal wounds is achieved after staining the cut walls with a solution of the chromophore indocyanine green, and irradiating them with a diode laser (810 nm) operating at low power (60 to 90 mW). The result is a localized heating of the cut, inducing controlled welding of the stromal collagen. In order to optimize this technique and to study the healing process, experimental tests, simulating cataract surgery and penetrating keratoplasty, were performed on rabbits: conventional and laser-induced suturing of corneal wounds were thus compared. A follow-up study 7 to 90 days after surgery was carried out by means of objective and histological examinations, in order to optimize the welding technique and to investigate the subsequent healing process. The analyses of the laser-welded corneas evidenced a faster and more effective restoration of the architecture of the stroma. No thermal damage of the welded stroma was detected, nor were there foreign body reactions or other inflammatory processes.

Laser tissue welding in ophthalmic surgery

Laser welding of ocular tissues is an alternative technique or adjunct to conventional suturing in ophthalmic surgery. It is based on the photothermal interaction of laser light with the main components of the extracellular matrix of connective tissues. The advantages of the welding procedure with respect to standard suturing and stapling are reduced operation times, lesser inflammation, faster healing and increased ability to induce tissue regeneration. The procedure we set up is based on the use of an infrared diode laser in association with the topical application of the chromophore Indocyanine Green. Laser light may be delivered either continuously or in pulses, thus identifying two different techniques that have been applied clinically in various types of transplants of the cornea.

Laser welding in penetrating keratoplasty and cataract surgery in pediatric patients: Early results

Purpose To evaluate the efficacy of diode laser welding to close corneal wounds in penetrating keratoplasty (PKP) and cataract surgery in pediatric patients. Setting Ophthalmology Department, Bambino Gesù Children’s Hospital, Rome, Italy. Design Prospective observational study. Methods Patients had surgery for congenital cataract (Group 1) or femtosecond laser–assisted PKP (Group 2). The surgery was followed by corneal wound closure using diode laser welding of the stroma. In Group 1, no standard suturing was used. In Group 2, the donor button was sutured onto the recipient using 8 single nylon sutures or a 10‐0 nylon running suture (12 passages). Laser welding was then used as an adjunct to the traditional suturing procedure. Results Group 1 comprised 7 eyes (7 patients; mean age 8.1 years ± 5.3 [SD], range 1 to 15 years) and Group 2, 5 eyes (5 patients; mean age 10.6 ± 3.3 years, range 6 to 15 years). The adhesion of the laser‐welded tissues was perfect; there were no collateral effects, and restoration of the treated tissues was optimum. Seidel testing showed no wound leakage during the follow‐up. Postoperative astigmatism did not change significantly from the first day after cataract surgery and shifted moderately 3 months after PKP. Conclusion Laser welding of corneal tissue appeared to be safe and effective in children for whom a sutureless surgical procedure is important to reduce the use of anesthesia for suture management, prevent endophthalmitis, and improve the antiamblyopic effect. Financial Disclosure No author has a financial or proprietary interest in any material or method mentioned.

First application of laser welding in clinical transplantation of the cornea

After a 4-year-long pre-clinical experimentation carried out at first on enucleated eyes and then on animal models, we applied a new procedure of laser welding of the cornea on voluntary patients. The welding technique is based on controlled irradiation of the cornea by diode laser radiation (805 nm) operating at low power (60-90 mW) in association with a photoenhancing chromophore applied locally. The welding effect is very effective and selective, because it takes place only in the cut where chromophore is present, while the contiguous tissue remains completely untouched. In the clinical phase, this technique was firstly tested in corneal cuts of increasing length on 25 patients subjected to facoemulsification of the cataract, by both sclero-corneal and corneal tunnels, and to extracapsular cataract extraction by sclero-corneal and corneal cuts. As previously confirmed by histological analysis performed on animal samples, we observed in humans too an early and effective healing process, with a sensible reduction of the post-operatory astigmatism. Based on these positive results, we finally arrived at the first application of diode laser- assisted corneal welding to penetrating keratoplasty (corneal transplantation), where this technique has been employed as far as now in 3 cases to substitute the application of the continuous suture.

A New Technique for the Closure of the Lens Capsule by Laser Welding

A new method is presented for the closure of the lens capsule based on laser welding of suitably prepared patches of anterior capsular tissue. Experiments were performed in freshly enucleated porcine eyes. The patches were previously stained with a solution of indocyanine green in sterile water and then welded on the recipient capsule by means of diode laser radiation at 810 nm. The welded tissue revealed mechanical properties comparable to those of healthy tissue. This technique is proposed to repair capsular breaks and to provide the closure of the capsulorhexis in lens refilling procedures.

Laser-Assisted Penetrating Keratoplasty: 1-Year Results in Patients Using a Laser-Welded Anvil-Profiled Graft

PURPOSE To describe a new laser-assisted penetrating keratoplasty technique combining a femtosecond anvil-like trephination pattern with the laser welding procedure. DESIGN Cohort prospective study with 1 year of follow-up (June 2011 through January 2013). METHODS This study was performed at Misericordia e Dolce Public Hospital, Prato, Italy. Twenty-four eyes of 22 patients underwent penetrating keratoplasty: 4 had granular dystrophy, 12 had keratoconus in its final stages, 3 had post-herpetic leukoma, and 5 had corneal scars. A femtosecond laser was used to create anvil-profiled cuts in donor and recipient corneas. Diode laser welding was performed, supporting standard suturing. All patients were evaluated for corrected distance visual acuity, pachymetry, manifest astigmatism, and endothelial cell density after 1, 3, 6, and 12 months. RESULTS Mean ± standard deviation postoperative corrected visual acuity was 0.48 ± 0.23 logarithm of the minimal angle of resolution (logMAR), 0.30 ± 0.18 logMAR, 0.18 ± 0.13 logMAR, and 0.13 ± 0.16 logMAR at 1, 3, 6, and 12 months, respectively. At the same follow-up times, mean pachymetry was 537 ± 57 μm, 533 ± 74 μm, 528 ± 72 μm, and 529 ± 58 μm, respectively; and mean endothelial cell density was 1945 ± 371 cells/mm(2), 1881 ± 410 cells/mm(2), 1781 ± 401 cells/mm(2), and 1730 ± 376 cells/mm(2), respectively. Mean manifest and topographic postoperative astigmatism were: 3.6 ± 2.5 diopters (D) and 4.65 ± 2.57 D at 1 month, 2.93 ± 2.34 D and 4.79 ± 2.85 D at 3 months, 2.82 ± 1.75 D and 3.44 ± 2.28 D at 6 months, and 2.08 ± 1.25 D and 2.73 ± 2.01 D at 12 months, respectively. All surgical operations were successful and without intraoperative complications. CONCLUSIONS The use of the anvil trephination profile was effective for performing laser-assisted penetrating keratoplasty. The large donor-recipient interface enables the laser welding procedure and good preservation of the recipients endothelial cell pool.

Preliminary experiences on diode laser welding in corneal transplantation

An experimental study on the applicability of diode laser welding to assist corneal transplantation is presented. Fusion of corneal tissue was accomplished by low power laser irradiation at 810 nm in association with Indocyanine Green (ICG) as the photoenhancing chromophore. Twenty freshly enucleated eyes obtained from young goats were used. In the preliminary experimental phase we examined the effects of laser welding on cuts produced by the scalpel on the cornea. Then we tried to design and test a novel diode laser- assisted corneal transplantation procedure. Our experimental results indicate that this procedure is affordable, but some technical aspects, as the application of ICG to the cut as well as the control of laser action are very critical issues.